Projects launched with the support of the ECRT

Connecting time-dependent degeneration to long-term tissue function in human Achilles tendon rupture

Delayed treatment of injured tissues, including tendons, has significant consequences for post-intervention healing and regeneration. Previous work in human Achilles tendon rupture (ATR) has shown the injured tissues exhibit higher degeneration when patients wait longer to receive surgical intervention. To date, it remains unclear how the status of the initial injured tissue affects healing, regeneration, and function of the healed Achilles tendon (AT). Current standards utilize ultrasonography to characterize AT function, but limited work has only been performed in ATR patients, and even fewer have investigated acute biological profiles with functional outcomes. The objective of our project is to connect long-term, post-ATR tendon function in humans with the time-dependent “healing profile” of the tissue at the time of treatment.  In this kickbox phase, we will assess in vivo AT function in patients that have received surgical treatment We anticipate that the results from this work will assist in the identification of possible targets for future, which will both improve post-injury tendon regeneration as well as functional patient outcomes.

Team: Alison Agres, Britt Wildemann, Sebastian Manegold, Kirsten Legerlotz

Contact: Alison Agres

You're stayin' alive, stayin' alive – Culturing human B cells in a 3D biomimetic environment

B cells are antibody-secreting lymphocytes at the center of the adaptive humoral immune system. They are challenging to culture, rendering it virtually impossible to follow their maturation path in vitro. This is mainly due to difficulties accessing their site of origin: the bone marrow (BM). Information on B cell development is mainly available from mouse and rat studies; however, in recent years it became clear that murine models are insufficient to mimic B cell biology in humans. As a team of synthetic biologists, immunologists, toxicologists, leukemia and stem cells experts, along with clinicians, we aim to develop a novel, reliable human BM system, allowing the study of B cells in a near in vivo environment.

If successful, the BM model could give completely new insights into B cell development in humans. In addition, the possibilities of translating it into therapeutic approaches, such as culturing B cells for clinical and commercial use, are immense, underscoring the benefits and applicability of our model in basic research.

Team: Melanie Ort, Christine Consentius, Alessandro Camponeschi, Anastasia Rakow, Sven Geißler, Janosch Schoon

Contact: Melanie Ort

A Quantitative Approach to ECM Function in Aged Muscle Regeneration – Towards Predictive Modeling of Cell-Matrix Interplay

Ageing affects the regenerative abilities of skeletal muscle resulting in compromised healing associated with fibrosis, impairing mobility and affecting quality of life. Current research is largely focused on intrinsic muscle stem cell function, but local extrinsic changes are mostly neglected. However, skeletal muscle repair relies on a dynamic interplay between muscle satellite cells (SCs) and the extracellular matrix (ECM) microenvironment. We have preliminary evidence for a transient developmental-like ECM during early phases of muscle regeneration. We believe that this creates a biomechanical threedimensional micro-niche conducive to SC expansion, differentiation and self-renewal during regeneration, and that its derailment in aging or disease contributes to SC malfunction. However the mechanical properties of the pro-regenerative ECM, especially during aging, has not been analyzed and the influence of the bio-mechanical properties of the aged ECM on stem cell behavior remains unclear. By combining experimental and mathematical analysis in an interdisciplinary approach, we aim to comparatively determine and quantify the dynamic spatio-temporal structure, composition and functionality of the transitory ECM in young and old mice. Iterative mathematical modeling via integration of biological data in a stepwise fashion will develop a predictive landscape to achieve a new level of understanding of ageing processes prospectively transferable to muscle-degenerative disorders.

Team: Sigmar Stricker, Max von Kleist, Arunima Murgai, Georgios Kotsaris, Vikram Sunkara

Contact: Arunima Murgai, Sigmar Stricker

3D - ALL - The tumor microenvironment of ALL in an bone marrow model

The aim of the project is to set up an as possible true-to-life model to study the microenvironment of primary  acute  lymphoblastic  leukemia  (ALL)  cells  on  cell  survival  and  resistance  development.  It  is planned to investigate the effect of different drugs especially on the microenvironment but also on the ALL cells. While most cancer research focuses mainly on the pharmaceutical impact on tumor cells, the approach of this project will concentrate on the bone marrow stroma. We plan to establish a co-culture of ALL cells with 3D cultured Mesenchymal Stroma Cells (MSCs), as published by Sieber et al. (2017). For that, a hydroxyapatit coated zirconium oxide ceramic is seeded with  patient  derived  MSCs  and  ALL  cells  to  mimic  the  in  vivo  conditions.  The  whole  cultivation  is performed  dynamically  in  the  "Multi-Organ-Chip"  (MOC).  The  MOC-platform,  which  has  been developed  at  the  chair  of  Medical  Biotechnology  at  Technical  University,  is  a  microfluidic  device consisting of a circular channel system which connects wells for the culture of different small functional human organ units, called organoids. Currently used 2D stroma cultures fail in their ability to support primary ALL cell proliferation and are therefor limited to test regenerative potentials of MSC cultures. This minimized model of ALL could be used for drug testing and dosage estimation. This in vitro leukemia model would present a breakthough for detailled studies of the human marrow and for personalized drug screenings.

Team: Kübrah Keskin, Tessa Skroblyn, Domenic Schlauch, Cornelia Eckert, Mark Rosowski

Contact: Kübrah Keskin

Alterations in endothelial cell mechanics affect nuclear morphology and chromatin conformation: the mechanogenomic code

J. Jatzlau/ P. Knaus in cooperation with Max Planck Institute for Molecular Genetics /S. Mundlos and M. Robson

Visualization of the endogenous BMP receptor ALK2 in endothelial cells using CRISPR/Cas9 and induced pluripotent stem cells

S. Hildebrandt/P. Knaus in cooperation with H. Stachelscheid Berlin Center for Regenerative Therapies

Serial high-resolution 3D-immunofluorescence bone imaging in patients undergoing hematopoietic stem cell Transplantation

I. Na/O. Penack Charité - Universitätsmedizin Berlin, Hematology, Oncology and Tumor Immunology

Systems Biology pipeline for discerning the disease phenotype of chondrocytes in osteoarthritis of the knee

V. Sunkara/A. Lang

Immunosuppressant-resistant anti-viral T cells for advanced adoptive T cell therapy

L. Amini/D. Wagner in cooperation with Institute for Medical and Human Genetics and Clinic for Nephrology and Internal Intensive Care

Tendon healing, scar formation or chronic inflammation – a matter of miscommunication between tenocytes and immune cells?

B. Sawitzki/B. Wildemann in cooperation with Department of Medicine for Gastroenterology and Rheumatology

Control of Human Tissue Homeostasis and Immunity by Helper T cells (TiSSueHeLP)

A. Thiel/K. Wolk in cooperation with Technische Universität Berlin

Expansion of SpCas9-specific regulatory T cells as an approach to prevent hazardous inflammatory damage to CRISPR/Cas9-edited tissues

M. Schmück-Henneresse in cooperation with Department of Pediatrics, Divison of Pneumology and Immunology